阿帕替尼对胃癌HGC-27细胞放射敏感性的影响及其作用机制

Effect of Apatinib on radiosensitivity of gastric cancer HGC-27 cells and its mechanism

目的 探讨阿帕替尼对胃癌HGC-27细胞放射敏感性的影响及可能的作用机制。方法 以不同浓度阿帕替尼(5μmol/L、10μmol/L、15μmol/L、20μmol/L)以及不同照射剂量(2 Gy、4 Gy、6 Gy、10 Gy、20 Gy)分别处理HGC-27细胞,再选取2 Gy和6 Gy单独照射或联合10μmol/L阿帕替尼处理HGC-27细胞;采用ELISA法检测细胞中血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达情况,CCK-8法检测细胞增殖能力,流式细胞术检测细胞凋亡和细胞周期,免疫荧光染色技术检测γ-H2AX的表达情况。结果 阿帕替尼呈浓度及时间依赖性抑制胃癌HGC-27细胞增殖(均P<0.05);不同剂量照射可促进细胞VEGF表达释放,且呈时间及剂量依赖性(均P<0.01)。10μmol/L阿帕替尼分别联合2 Gy和6 Gy照射后,HGC-27细胞增殖抑制作用、细胞凋亡率及G2/M期的细胞比例均较单照组升高(均P<0.01),且6 Gy联合组的作用强度大于2 Gy联合组(均P<0.01)。6 Gy联合组细胞核内γ-H2AX焦点淬灭较6 Gy单照组延迟。结论 阿帕替尼通过抑制细胞增殖,促进细胞凋亡并诱导细胞周期再分布增强胃癌HGC-27细胞放射敏感性,其作用机制可能与延迟γ-H2AX表达而干扰DNA双链断裂修复有关。

ObjectiveTo investigate the effect of Apatinib on the radiosensitivity of gastric cancer HGC-27 cells and its possible mechanism.MethodsHGC-27 cells were treated with different concentrations of Apatinib(5 μmol/L,10 μmol/L,15 μmol/L,20 μmol/L) and different irradiation doses(2 Gy,4 Gy,6 Gy,10 Gy,20 Gy).HGC-27 cells were then irradiated with 2 Gy and 6 Gy alone or combined with 10 μmol/L Apatinib.The expression of vascular endothelial growth factor(VEGF)was detected by ELISA,cell proliferation capacity was detected by CCK-8,cell apoptosis and cell cycle were detected by flow cytometry,and expression of γ-H2AX was detected by immunofluorescence staining.ResultsApatinib inhibited the proliferation of gastric cancer HGC-27 cells in a concentrationand time-dependent manner(all P<0.05).Different doses of irradiation promoted the release of VEGF in a time-and dose-dependent manner(all P<0.01).After the combination of 10 μmol/L Apatinib with 2 Gy and 6 Gy irradiation,respectively,the inhibition of cell proliferation,apoptosis rate and proportion of cells in G2/M phase of HGC-27 cells were increased compared with those in the single irradiation group(all P<0.01).The effect intensity of 6 Gy combined group was greater than that of 2 Gy combined group(all P<0.01).The nucleus focal quenching of γ-H2AX in the 6 Gy combined group was delayed compared with that in the 6 Gy single irradiation group.ConclusionsApatinib enhances the radiosensitivity of gastric cancer HGC-27 cells by inhibiting cell proliferation,promoting cell apoptosis and inducing cell cycle redistribution.The underlying mechanism may be related to the delay of γ-H2AX expression and the interference of DNA double-strand fracture repair.